KR100288117B1 - Cacao extract including dietary fiber - Google Patents
Cacao extract including dietary fiber Download PDFInfo
- Publication number
- KR100288117B1 KR100288117B1 KR1019990013401A KR19990013401A KR100288117B1 KR 100288117 B1 KR100288117 B1 KR 100288117B1 KR 1019990013401 A KR1019990013401 A KR 1019990013401A KR 19990013401 A KR19990013401 A KR 19990013401A KR 100288117 B1 KR100288117 B1 KR 100288117B1
- Authority
- KR
- South Korea
- Prior art keywords
- cacao
- dietary fiber
- extract
- weight
- water
- Prior art date
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23G1/00—Cocoa; Cocoa products, e.g. chocolate; Substitutes therefor
- A23G1/30—Cocoa products, e.g. chocolate; Substitutes therefor
- A23G1/32—Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds
- A23G1/48—Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds containing plants or parts thereof, e.g. fruits, seeds, extracts
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23G1/04—Apparatus specially adapted for manufacture or treatment of cocoa or cocoa products
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23G1/30—Cocoa products, e.g. chocolate; Substitutes therefor
- A23G1/32—Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds
- A23G1/42—Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
- A23G1/426—Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing vitamins, antibiotics
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G1/00—Cocoa; Cocoa products, e.g. chocolate; Substitutes therefor
- A23G1/30—Cocoa products, e.g. chocolate; Substitutes therefor
- A23G1/56—Cocoa products, e.g. chocolate; Substitutes therefor making liquid products, e.g. for making chocolate milk drinks and the products for their preparation, pastes for spreading, milk crumb
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
- A23L33/22—Comminuted fibrous parts of plants, e.g. bagasse or pulp
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A23G2200/00—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF containing organic compounds, e.g. synthetic flavouring agents
- A23G2200/14—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF containing organic compounds, e.g. synthetic flavouring agents containing fruits, nuts, e.g. almonds, seeds, plants, plant extracts, essential oils
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Abstract
본 발명은 식이섬유를 함유한 카카오 추출물에 관한 것으로서, 더욱 상세하게는 종래의 카카오(Cacao)의 원료콩으로부터 카카오 열매만 사용하고 폐기되던 카카오 콩 껍질을 용매추출하여 잔사로 얻는 추출물로써, 여기에는 식이섬유가 과량 함유되어 있으며, 특히 불용성 식이섬유가 수용성 식이섬유 보다 과량 함유되어 있고 이들의 보수력, 보유능, 점도특성, 글루코오스 및 담즙산 투과 저지율의 물리 화학적 특성이 우수할 뿐만 아니라, 혈당 강하 효과 및 콜레스테롤 대사 개선 효과 등 장질환과 대사성 질환의 생리활성을 향상시키므로서 당뇨병의 치료에 유용한 식이섬유를 함유한 카카오 추출물에 관한 것이다.The present invention relates to a cacao extract containing dietary fiber, and more specifically, using only cacao fruit from the cacao raw beans of the conventional cacao extract extract obtained by solvent extraction of the discarded cacao beans peel as a residue, It contains excessive amounts of dietary fiber, especially insoluble dietary fiber in excess of water-soluble dietary fiber, its water holding capacity, retention capacity, viscosity characteristics, physicochemical properties of glucose and bile acid permeation inhibition rate, as well as hypoglycemic effect and cholesterol It relates to cacao extract containing dietary fiber useful for the treatment of diabetes by improving the physiological activity of intestinal diseases and metabolic diseases such as metabolic improving effect.
Description
본 발명은 식이섬유를 함유한 카카오 추출물에 관한 것으로서, 더욱 상세하게는 종래의 카카오(Cacao)의 원료콩으로부터 카카오 열매만 사용하고 폐기되던 카카오 콩 껍질을 용매추출하여 잔사로 얻는 추출물로써, 여기에는 식이섬유가 과량 함유되어 있으며, 특히 불용성 식이섬유가 수용성 식이섬유 보다 과량 함유되어 있고 이들의 보수력 , 보유능, 점도특성, 글루코오스 및 담즙산 투과 저지율의 물리 화학적 특성이 우수할 뿐만 아니라, 혈당 강하 효과 및 콜레스테롤 대사 개선 효과 등 장질환과 대사성 질환의 생리활성을 향상시키므로서 당뇨병의 치료에 유용한 식이섬유를 함유한 카카오 추출물에 관한 것이다.The present invention relates to a cacao extract containing dietary fiber, and more specifically, using only cacao fruit from the cacao raw beans of the conventional cacao extract extract obtained by solvent extraction of the discarded cacao beans peel as a residue, It contains excessive amounts of dietary fiber, especially insoluble dietary fiber in excess of water-soluble dietary fiber, and their physicochemical properties of water retention, retention capacity, viscosity characteristics, glucose and bile acid permeation inhibition rate, as well as hypoglycemic effect and cholesterol It relates to cacao extract containing dietary fiber useful for the treatment of diabetes by improving the physiological activity of intestinal diseases and metabolic diseases such as metabolic improving effect.
카카오(Theobroma cacao L.)는 라틴 아메리카 원산의 벽오동나무(Byttneriaceae)과에 속하는 다년생 식물로서, 벽오동 나무의 높이는 6 ∼ 8 m까지 자란다. 이것은 약 15 ∼ 30 ㎝크기의 달걀모양(oval) 형태의 열매(pod)를 맺게 되는데, 이 열매 안에는 약 30 ∼ 40 개의 종자(seed)가 과육(pulp)으로 둘러싸여 있다. 카카오 열매는 껍질(shell 또는 testa), 내부 열매(nib 또는 cotyledon) 및 배아(germ)로 구성되어 있으며, 이중에서 실제 초콜렛의 주원료로 사용되는 부분은 버터 함유량이 많은 니브(nib)를 갈아서 만든 분쇄물이다. 이러한 분쇄물을 카카오 분비액(cacao liquor) 또는 카카오 매스(cacao mass, CM)라고 부르는데, 분쇄물이 버터의 융점이하에서는 페이스트상을 나타내기 때문이다.Cacao (Theobroma cacao L.) is a perennial plant belonging to the family of the paulownia (Byttneriaceae) native to Latin America, the height of the paulownia tree grows up to 6-8 m. It forms an egg-shaped pod of about 15 to 30 cm in size, with about 30 to 40 seeds surrounded by a pulp. Cacao is composed of shell (testa), inner fruit (nib or cotyledon), and germ, and the part that is used as the main raw material of chocolate is crushed by grinding nib with high butter content Water. Such a pulverized product is called a cacao liquor or a cacao mass (CM) because the pulverized product has a paste form below the melting point of the butter.
이와 같이 내부 열매는 초콜렛의 주원료로 사용되는 반면, 카카오 매스 생산 공정 중에 탈각되는 껍질은 약 15 중량%(전세계적으로 약 40만 M/T)의 부산물로서 산출되어 전량 폐기되고 있으며, 이것을 카카오 콩 껍질(cacao bean husk, CBH)이라고 한다.As such, the inner fruit is used as the main raw material of chocolate, while the shell shelled during the cacao mass production process is calculated as a by-product of about 15% by weight (about 400,000 M / T worldwide), which is entirely discarded. It is called the cacao bean husk (CBH).
한편, 현재의 국내 초콜렛 시장은 매우 큰 시장을 형성하고 있지만, 최근에 와서는 외국산 초콜렛의 수입 개방과 함께 초콜렛에 대한 이미지가 달라지고 있다. 특히, 최근의 다이어트 붐이나 당뇨, 충치, 동맥경화 등 각종 장질환 및 대사성 질환과 관련한 건강 식품에 대한 소비자 관심의 고조로 당함량이 높으며 고지방 열량식의 인식을 갖게된 초콜렛에 대한 부정적 인식은 점차 커지고 있는 실정이다.On the other hand, the current domestic chocolate market forms a very large market, but in recent years, the image of chocolate is changing with the opening of imports of foreign chocolate. In particular, the recent increase in dietary boom, diabetes, tooth decay, arteriosclerosis, and other foods related to health problems related to various intestinal and metabolic diseases, such as high sugar content and high-fat calorie, the negative recognition of chocolate gradually It is growing.
따라서, 이에 대한 대응방안으로 초콜렛의 새로운 생리 기능성을 규명하여 건강 식품으로 인식하는 것이 요구되는 바, 최근 초콜렛의 주성분인 카카오 매스 및 카카오 매스 생산공정 중에서 탈각되던 가공 폐기물인 카카오 콩 껍질에 함유된 폴리페놀류가 각종의 생리활성을 나타내어 지난 몇 년 동안 일본을 중심으로 연구되고 있다. 이러한 생리기능성 연구에 대하여 참고문헌['Isolation of Antioxidants from Cacao Bean Husk and its Application', New Food Industry, 26,1,1984]에 공지되어 있으며, 특히 카카오 열매나 카카오 매스에 들어있는 폴리페놀류 성분들이 동맥경화, 알레르기 질환 및 각종 심장질환에 대한 새로운 면역 조절작용을 보이므로 암의 예방 효과, 항충치, 항산화, 항스트레스 작용 등의 다양한 생리기능을 갖는 것으로 발견되어 크게 주목 받고있다. 이에 따라 국내에서도 그 동안 초콜렛의 주성분인 카카오 매스는 물론, 전량 폐기되었던 카카오 콩 껍질에 대한 연구를 진행하고 있으며, 이들 성분으로부터 기인되는 지타제(GTase, Glycosyltransferase), ACE(Angiotesin coverting enzyme), 엑소아제(Xoase, Xanthine oxidase) 및 티로시나제(tyrosynase) 효소에 대한 저해제로서의 작용 등의 각종 생리활성을 밝힘으로써, 초콜렛 관련 성분에 대한 새로운 기능성 탐색에 큰 성과를 거두고 있다['Cacao Bean Husk 추출물로부터 분리한 충치예방 물질의 구조결정', 한국생물공학회지, 8(1), 69 ∼ 74, 1993].Therefore, as a countermeasure, it is required to identify new physiological functionalities of chocolate and recognize them as health foods. Phenols exhibit various biological activities and have been studied in Japan for the past several years. This physiological function study is known in the reference 'Isolation of Antioxidants from Cacao Bean Husk and its Application', New Food Industry, 26,1,1984. In particular, polyphenols contained in cacao or cacao mass As it shows new immunomodulatory effects on atherosclerosis, allergic diseases and various heart diseases, it has been found to have various physiological functions such as cancer prevention effect, anti tooth decay, antioxidant, and antistress action. Accordingly, in Korea, we are conducting research on cacao beans, the main ingredient of chocolate, as well as cacao beans, which have been completely discarded.These are derived from these enzymes: GTase (Glycosyltransferase), ACE (Angiotesin coverting enzyme), and exo. By identifying various physiological activities such as inhibitors against enzymes (Xoase, Xanthine oxidase) and tyrosynase enzymes, the researchers have achieved great results in the search for new functionalities for chocolate-related components [Isolated from 'Cacao Bean Husk Extract Structural Determination of Caries Prevention Materials', Journal of Biotechnology and Biotechnology, 8 (1), 69-74, 1993].
상기에서 언급한 각종 생리 기능성과 관련하여 가장 주목받고 있는 또 다른 소재의 하나로는 카카오 콩에 풍부하게 함유되어 있는 식이섬유(Dietary fiber)가 있는데 이에 관련한 연구가 참고문헌['Dietary Fiber', Food Technol., 43, 133, 1989, 'Dietary Fiber and Desease', J. Am. Med. Assoc., 229, 1068, 1974]에 공지되어 있다. 그러나, 식이섬유는 사람의 소화 효소로는 소화되지 않는 식물 세포의 구조 잔사로서, 소화 및 흡수가 잘 되지 않기 때문에 그 동안 영양적 가치가 없는 것으로 인식되었고, 식이섬유는 카카오 열매나 카카오 콩 껍질에도 매우 풍부하게 함유되어 있지만, 카카오 식이섬유의 소재 특성 및 생리적 기능성에 대해서는 폴리페놀류에 비해서 연구가 미흡하다. 그러나, 1972년 트로웰(Trowell)의 보고에 의하면 식이섬유의 섭취 부족이 비만증, 심장계 질환, 당뇨병, 장 질환 및 담석과 같은 질병들과 관련된다고 하였으며, 그 이후 식이섬유에 대한 생리 기능성 연구와 관심['Definition fo Dietary Fiber and Hypotheses That It is a Protective Factor in Certain Diseases', Am. J. Clin. Nutr., 29, 417, 1976, 'Metabolic Effects of Dietary Pectins Related to Human Health', Food Technol., 41, 91, 1987]이 크게 고조되고 있다.Another material that is attracting much attention in relation to the various physiological functions mentioned above is dietary fiber, which is rich in cacao beans, and related research is referred to as 'Dietary Fiber', Food Technol. ., 43, 133, 1989, Dietary Fiber and Desease, J. Am. Med. Assoc., 229, 1068, 1974. However, dietary fiber is a structural residue of plant cells that is not digested by human digestive enzymes, and has been recognized as having no nutritional value because of poor digestion and absorption. Although it is very rich in content, the material properties and physiological functionalities of cacao dietary fiber are less studied than polyphenols. However, in 1972, Trowell reported that a lack of dietary fiber was associated with diseases such as obesity, heart disease, diabetes, intestinal disease and gallstones. '' Definition fo Dietary Fiber and Hypotheses That It is a Protective Factor in Certain Diseases', Am. J. Clin. Nutr., 29, 417, 1976, Metabolic Effects of Dietary Pectins Related to Human Health, Food Technol., 41, 91, 1987.
통상의 식이섬유는 여러 가지 식품소재에 함유되어 있으며 그 종류도 대단히 많다. 이러한 식이섬유는 크게 수용성 식이섬유(SDF, soluble dietary fiber)와 불용성 식이섬유(IDF, insoluble dietary fiber)로 나눌 수 있다. 수용성 식이섬유는 난소화성 덱스트린, 풀루란, 폴리덱스트로오스, 글루코 만난, 펙틴, 구아껌 효소 분해물, 미강, 헤미 셀룰로오스 및 한천 등에 많이 함유되어 있으며, 불용성 식이섬유는 밀기울, 스피룰리나, 키토산, 옥수수, 사과, 대두, 대두껍질, 셀룰로오스 및 인삼 등에 풍부하게 함유되어 있다. 각종 식물성 식품재료의 식이섬유 함량은 그 종류에 따라 1 ∼ 43 중량% 범위로 존재한다.Common dietary fiber is contained in many food materials, and there are many kinds. These dietary fibers can be largely divided into soluble dietary fiber (SDF) and insoluble dietary fiber (IDF). Water soluble fiber is high in indigestible dextrin, pullulan, polydextrose, glucomannan, pectin, guam gum degradant, rice bran, hemi cellulose and agar, and insoluble fiber is bran, spirulina, chitosan, corn, Rich in apples, soybeans, soybean hulls, cellulose and ginseng. The dietary fiber content of various plant food materials is present in the range of 1 to 43% by weight, depending on the type.
또한, 식이섬유의 생리활성은 물리 화학적 특성인 보수력, 양이온 교환능, 담즙산 결합력, 발효성 등에 영향을 받으며 식이섬유의 구성성분과 조성, 함량, 결합상태, 입자크기, 식품의 종류, 가공법, 조리조건 및 식이섬유원 등에 따라 그 기능이 다르게 나타난다['식이섬유의 생리활성과 이용', 식품과학과 산업, 28(3), 1995, 'The Effect of the Polysacchride Composition and Structure of Dietary Fiber on Cecal Fermentation and Fecal Excretion', Am. Soc. Clin. Nutr., 51 ∼ 55, 1986]. 일반적으로 식이섬유원의 수분 흡착력은 소화력의 저하, 변의 부피와 무게 증가, 혈청 중성지질(triglyceride)을 낮추는 기전과 관련이 있는 것으로 알려지고 있다. 특히, 곡류섬유의 물 흡수능은 변용적 증가효과의 중요 결정인자로 알려지고 있다. 이러한 수분 흡착력은 식이섬유의 종류, 함량, 입자크기에 따라 크게 영향을 받으며 주로 식이섬유의 구성성분, 입도, pH 및 이온강도에 따라서 좌우된다.In addition, the physiological activity of dietary fiber is affected by physicochemical properties such as water retention, cation exchange capacity, bile acid binding ability, fermentation, etc., and the composition, content, binding state, particle size, food type, processing method, and cooking conditions of dietary fiber. And its function varies according to dietary fiber sources ['physiological activity and use of dietary fiber', Food Science and Industry, 28 (3), 1995, 'The Effect of the Polysacchride Composition and Structure of Dietary Fiber on Cecal Fermentation and Fecal'. Excretion ', Am. Soc. Clin. Nutr., 51-55, 1986]. In general, the water adsorption capacity of dietary fiber sources is known to be related to the mechanism of lowering digestion, increasing stool volume and weight, and lowering serum triglyceride. In particular, the water absorption capacity of grain fiber is known as an important determinant of variability increase effect. This moisture adsorption capacity is greatly affected by the type, content, and particle size of the fiber, and mainly depends on the composition, particle size, pH and ionic strength of the fiber.
한편, 당뇨병과 관련하여 여러 물질들에 대한 연구가 진행되고 있는데, 이에 대한 치료법으로는 식이요법, 운동요법 및 약물치료 등이 사용되고 있지만, 이중 약물치료에서는 독성 및 환자의 내성 문제가 대두되고 있기 때문에 점차 천연물질에서의 새로운 약리물질을 탐색하는 연구['당뇨병', 전남대학교 출판부, 1990]가 널리 진행되고 있다.On the other hand, studies on various substances related to diabetes are being conducted. As a treatment method, diet, exercise therapy, and drug treatment are used, but of these drugs, toxicity and patient tolerance problems are emerging. Increasingly, researches on new pharmacological substances in natural materials ['Diabetes', Chonnam National University Press, 1990] are being widely conducted.
천연물질 중에서 카카오 유래 식이섬유에 대한 당뇨병의 혈압 강하나 콜레스테롤의 저하에 관한 연구는 일부 보고['초콜렛, 코코아의 최신 연구동향', 제 1회 초콜렛 & 카카오 국제학술 심포지엄 논문집, 9 ∼ 20, 1998]된 바 있으나, 실용화되지 못하고 있으며 지속적인 연구가 저조한 실정이다. 특히, 각종 식이 섬유원에서 널리 보여지는 당뇨에 대한 효과 및 물성 기능에 관한 보고는 아직 발표된 바 없으며, 일반적으로 식이섬유는 포도당의 흡수를 지연시켜 인슐린의 요구량을 감소시킴으로써, 당뇨병의 치료에도 도움이 되고 있는 것으로 알려져 있다['The effect of Indigestible Dextrin on Sugar Tolernce. I. Studies on Digestion Absorption and Sugar Tolerence[Japanese]', Nippon Naibunpi Gakkai Zasshi Folia Endocrinologica Japonica, 68, 623, 1992].Among the natural substances, a study on the blood pressure and cholesterol lowering of diabetes mellitus for cacao-derived fiber has been reported ['Chocolate, Cocoa's latest research trend', 1st Chocolate & Cacao International Symposium, 9-20, 1998]. Although it has been done, it has not been put into practical use and the research is poor. In particular, reports on the effects and physical functions on diabetes, which are widely found in various dietary fiber sources, have not been published. In general, dietary fiber delays the absorption of glucose and reduces the amount of insulin, thereby helping to treat diabetes. ['The effect of Indigestible Dextrin on Sugar Tolernce.' I. Studies on Digestion Absorption and Sugar Tolerence [Japanese] ', Nippon Naibunpi Gakkai Zasshi Folia Endocrinologica Japonica, 68, 623, 1992].
실제로 임상 연구에 의한 고식이섬유의 식이는 제Ⅱ형 당뇨병 환자의 내당능(glucose tolerance)을 개선하고 인슐린의 요구량을 유의적으로 저하시키는 것으로 밝혀졌다['The Prolonged Effect of a Low Cholesterol High Carbohydrate upon the Serum Lipids in Diabetic Patients', Diabetes, 12, 127 ∼ 132, 1963]. 아울러 당뇨병의 여러 합병증이 주로 고혈당증(hyperglycemia)과 직접 관계하므로 식후 고혈당증을 감소시키는 것은 중요한데, 적절한 식이섬유가 함유된 식사의 장기간 사용은 제Ⅱ형 당뇨병 환자의 총 혈청 콜레스테롤(total serum cholesterol) 수준을 유의적으로 낮춘다고도 보고되고 있다['Effect of a Fat Free, High Carbohydrate Diet on Diabetic Subjects with Fasting Hyperglycemia', Diabetes, 23, 138 ∼ 142, 1974, '한국인의 주식의 종류와 식이형태가 정상인과 제Ⅱ형 당뇨병 환자의 혈청 글루코오스, 인슐린 및 C-펩티드에 미치는 영향에 관한 연구', 숙명여자대학교 석사학위논문, 1987].Indeed, clinical studies have shown that dietary fiber improves glucose tolerance and significantly lowers insulin demand in patients with type II diabetes ['The Prolonged Effect of a Low Cholesterol High Carbohydrate upon the Serum Lipids in Diabetic Patients', Diabetes, 12, 127-132, 1963]. In addition, it is important to reduce postprandial hyperglycemia because many complications of diabetes are directly related to hyperglycemia. Long-term use of a diet containing adequate dietary fiber may reduce total serum cholesterol levels in patients with type II diabetes. It is reported to be significantly lowered ['Effect of a Fat Free, High Carbohydrate Diet on Diabetic Subjects with Fasting Hyperglycemia', Diabetes, 23, 138-142, 1974, A Study on the Effect of Serum Glucose, Insulin and C-Peptide in Patients with Type II Diabetes', Master's Thesis, Sookmyung Women's University, 1987].
이와 같이, 식이섬유에 대한 연구가 진행되고는 있지만 당뇨병 치료에 유의성이 있는 효과를 나타내는 천연 추출물의 개발은 아직 알려지지 않고 있으며, 특히 카카오 추출물이 특정 효능이나 인체의 질병치료에 미치는 영향에 대해서는 미흡한 실정이고, 그 연구도 매우 원론적 수준밖에 되지 못하고 있으며 실제로 그 동안 천연 식물에서 추출한 식이섬유의 당뇨병 치료의 적용과 제품화가 이루어지지 못하고 있었다.As such, although research on dietary fiber is ongoing, the development of natural extracts that have a significant effect on the treatment of diabetes is not known, and in particular, the effect of cacao extract on the specific efficacy and treatment of diseases of the human body is insufficient. In addition, the research is only at a very basic level, and in fact, the application and commercialization of diabetes treatment of dietary fiber extracted from natural plants has not been accomplished.
이에, 본 발명은 종래에 초콜렛의 주원료인 카카오 매스 가공 중 전량 폐기되었던 카카오 콩 껍질을 중요한 유용 자원으로 활용하기 위하여, 카카오 콩 껍질의 추출 잔사에서 식이섬유를 분리하여 소재 특성과 생리적 기능을 연구한 결과, 글루코오스와 담즙산 흡수 지연 효과, 혈당 강하 효과 및 콜레스테롤 대사가 개선되어 당뇨병의 치료에 유용한 식이섬유가 그 어느 식물보다 과량 함유되어 있다는 놀라운 사실을 알게되어 본 발명을 완성하였다. 따라서, 본 발명은 종래 카카오 가공 공정에서 폐기되었던 카카오 콩 껍질로부터 추출한 당뇨병의 치료 효과에 우수한 식이섬유를 함유한 카카오 추출물을 제공하는 데 그 목적이 있다.Thus, the present invention is to study the material properties and physiological functions by separating the dietary fiber from the extract residue of cacao bean husk in order to utilize the cacao bean husk that was discarded entirely during the processing of cacao mass, the main raw material of chocolate as an important useful resource As a result, the glucose and bile acid absorption delaying effect, the hypoglycemic effect and cholesterol metabolism have been improved to realize the surprising fact that the excess amount of dietary fiber useful for the treatment of diabetes than any other plant to complete the present invention. Accordingly, an object of the present invention is to provide a cacao extract containing dietary fiber excellent in the therapeutic effect of diabetes mellitus extracted from the cacao bean husk that was discarded in the conventional cacao processing process.
도 1은 L-BTC 공법에 의한 카카오 콩 껍질의 추출 잔사를 제조하는 공정을 나타낸 것이고,1 shows a process for preparing the extraction residue of cacao beans peel by the L-BTC method,
도 2는 식이섬유의 함량 확인을 위한 프로스키(Proskey) 법을 나타낸 것이고,Figure 2 shows the Proskey (Proskey) method for checking the content of dietary fiber,
도 3은 카카오 콩에서 추출한 각 성분의 식이 섬유 함량을 나타낸 것이고,Figure 3 shows the dietary fiber content of each component extracted from cacao beans,
도 4는 카카오 식이섬유의 글루코오스의 경시변화와 흡수 억제지수를 나타낸 것이며,Figure 4 shows the change over time and the absorption index of glucose of cacao dietary fiber,
도 5는 카카오 식이섬유의 담즙산의 경시변화와 흡수 억제지수를 나타낸 것이며,Figure 5 shows the changes over time and the absorption inhibition index of bile acids of cacao dietary fiber,
도 6은 카카오 식이섬유의 총 당화 혈색소를 나타낸 것이다.Figure 6 shows the total glycated hemoglobin of cacao dietary fiber.
본 발명에 있어서, 카카오 가공공정에서 폐기되는 카카오 콩 껍질의 추출 잔사로 얻어지며 총 식이섬유의 함량이 43 ∼ 60 중량%인 식이섬유를 함유한 카카오 추출물을 그 특징으로 한다.In the present invention, the cacao extract is obtained as an extract residue of the cacao bean husk discarded in the cacao processing process and contains a dietary fiber having a total dietary fiber content of 43 to 60% by weight.
이와 같은, 본 발명을 더욱 상세히 설명하면 다음과 같다.As described above, the present invention will be described in more detail.
본 발명은 카카오 원료콩으로부터 카카오 매스를 가공할 때, 전량 폐기되었던 카카오 콩 껍질을 용매추출하여 얻은 잔사에서 분리된 식이섬유를 함유하는 카카오 추출물로써, 특히 식이섬유가 카카오 열매나 기타 다른 식물에서보다 과량 함유되어 있어서 당뇨병의 혈당 강하와 콜레스테롤 대사 개선 효과에 우수한 생리활성을 나타내므로 당뇨병의 치료에 탁월한 효능이 있는 식이섬유를 함유한 카카오 추출물에 관한 것이다.The present invention is a cacao extract containing dietary fiber separated from the residue obtained by solvent extraction of the cacao bean husk, which has been completely discarded when processing cacao mass from cacao raw soybeans. The present invention relates to a cacao extract containing dietary fiber having an excellent effect on the treatment of diabetes mellitus because it contains an excess of physiological activity in diabetic hypoglycemic and cholesterol metabolism improving effect.
본 발명의 카카오 추출물은 카카오 원료콩을 예열, 분쇄, 선별 후 탈각하여 카카오 콩 껍질을 얻은 다음, 카카오 콩 껍질에 에탄올을 첨가 후 추출하여 잔사로 얻을 수 있다.The cacao extract of the present invention can be obtained by preheating, crushing and screening the cacao raw soybeans to obtain a cacao bean shell and then extracting after adding ethanol to the cacao bean husk as a residue.
본 발명에 있어서 카카오 콩 껍질의 추출 잔사를 제조하는 공정은 도 1과 같은 L-BTC(Better Taste and Color) 공법을 이용하여 제조할 수 있는데, 이와 같은 제조방법의 일례를 상세히 설명하면 다음과 같다.In the present invention, the process of preparing the extraction residue of the cacao bean husk can be prepared by using the L-BTC (Better Taste and Color) method as shown in Figure 1, when an example of such a manufacturing method will be described in detail as follows. .
우선, 카카오 원료콩을 0.5 ∼ 1 bar의 압력과 160 ∼ 180 ℃의 온도에서 열적 예열 처리한 다음 이것을 분쇄하고 선별하면, 카카오 원료콩의 껍질이 탈각되어 본 발명이 추출하고자 하는 원료 물질인 카카오 콩 껍질을 얻게 된다. 이때 압력이 0.5 bar 미만이면 니브중에 콩껍질이 과다하게 되는 문제가 있고, 1 bar를 초과하면 니브 수율(%)이 낮아지는 문제가 있어서 바람직하지 않다. 그리고, 상기 카카오 원료콩을 분쇄하기 전에 원료콩을 부드럽고 분쇄에 용이하도록 예열하는데, 예열시의 온도가 160 ℃보다 낮으면 콩껍질과 니브의 분리가 잘되지 않는 문제가 있고, 180℃보다 높으면 니브 수율이 떨어지는 문제를 초래한다.First, when the cacao raw soybean is thermally preheated at a pressure of 0.5 to 1 bar and at a temperature of 160 to 180 ° C., then pulverized and screened, the cacao soybean is shelled and the cacao soybean which is a raw material to be extracted according to the present invention. You get a shell. In this case, if the pressure is less than 0.5 bar, there is a problem that the bean shell in the nib is excessive, and if it exceeds 1 bar, there is a problem that the nib yield (%) is lowered, which is not preferable. And, before grinding the cacao raw beans, the raw beans are preheated to be soft and easy to grind. If the temperature at the time of preheating is lower than 160 ° C., there is a problem in that the separation of the bean husk and nib is not good. This leads to a problem of poor yield.
본 발명에 있어서, 상기의 예열 가공된 카카오 콩 껍질에 에탄올을 첨가하여 원심분리기로 추출하면 카카오 콩 껍질의 추출 잔사를 얻을 수 있다. 이 과정에서 폴리페놀성분이 제거되는데, 이 성분은 폴리페놀함유 카카오 음료의 제조에 사용하기 위해서이다. 이때, 추출용매는 에탄올, 메탄올, 부탄올 등이 사용되는데, 바람직하기로는 에탄올을 사용한다.In the present invention, ethanol is added to the preheated cacao bean husk and extracted with a centrifuge to obtain an extraction residue of cacao bean husk. This process removes the polyphenol component, which is intended for use in the production of polyphenol-containing cacao beverages. At this time, the extraction solvent is used ethanol, methanol, butanol and the like, preferably ethanol.
본 발명은 잔사의 식이섬유의 함량 확인을 위해 도 2와 같은 프로스키(Proskey) 법으로 식이섬유 성분을 분리할 수 있는데, 이 잔사에 테마밀(temamyl), 프로테아제(protease), 아밀로글루코시다제(amyloglucosidase)의 효소를 첨가해 본 결과, 본 발명의 카카오 추출물은 수용성 식이섬유와 불용성 식이섬유가 혼재된 바람직한 상태임이 확인된다.The present invention can separate the dietary fiber components by the Proskey (Proskey) method as shown in Figure 2 to determine the content of dietary fiber of the residue, this residue (temamyl), protease (amyloglucosid) As a result of the addition of the enzyme (amyloglucosidase), it was confirmed that the cacao extract of the present invention is in a preferable state in which water-soluble fiber and insoluble fiber are mixed.
한편, 본 발명의 카카오의 제조 공정에서 얻어질 수 있는 또 다른 물질로는 분쇄 및 선별하여 껍질이 제거된 카카오 분쇄물(winnowed nibs)과 1 ∼ 1.5 bar의 압력에서 10 ∼ 15분 동안 카카오 분쇄물을 반응 처리한 후 얻을 수 있는 카카오 니브(reacted nibs)가 있다. 그리고, 130 ∼ 140 ℃의 온도와 D.R.(Discharge Rate)이 900 ∼ 1100 ㎏/hr의 조건에서 건조 및 볶음 처리하고 갈아서 초콜렛의 주원료인 카카오 매스를 얻을 수 있다.On the other hand, another material that can be obtained in the manufacturing process of the cacao of the present invention is a cacao pulverized (winnowed nibs) by grinding and screening and cacao pulverized for 10 to 15 minutes at a pressure of 1 to 1.5 bar There are cacao nibs (reacted nibs) that can be obtained after the reaction treatment. And the cacao mass which is a main raw material of chocolate can be obtained by drying, roasting, and grinding under the conditions of 130-140 degreeC temperature and D.R. (Discharge Rate) 900-1100 kg / hr.
본 발명은 상기 추출물에 카카오 분쇄물, 카카오 니브, 카카오 매스 또는 이들의 혼합물 5 ∼ 15 중량%를 추가로 혼합할 수 있다.The present invention may further mix 5 to 15% by weight of cacao powder, cacao nib, cacao mass or a mixture thereof.
이와 같은 방법으로 제조된 본 발명에 따른 카카오 콩 껍질, 분쇄물, 니브, 매스의 탈지물의 총 식이섬유의 함량은 30 ∼ 60 중량%, 바람직하게는 43 ∼ 60 중량%로서, 종래의 1 ∼ 40 중량%보다 식이섬유가 과량 함유되어 있어서, 글루코오스 및 담즙산의 흡수지연 효과등의 생리활성이 매우 우수하다.The total dietary fiber content of the decay of cacao bean husk, ground product, nib, and mass prepared according to the present invention is 30 to 60% by weight, preferably 43 to 60% by weight. Since the dietary fiber is contained more than the weight%, the physiological activity such as absorption delay effect of glucose and bile acid is very excellent.
본 발명의 카카오 추출물인 카카오 콩 껍질의 에탄올 추출로 폴리페놀 성분이 제거된 잔사에 대한 불용성 식이섬유 함량은 도 3에서 알 수 있듯이 카카오 콩 껍질보다 증가한다. 이에 비해 상등액의 불용성 및 수용성 식이섬유 함량이 각각 0.2 ∼ 0.4 중량%와 2 ∼ 3 중량%로 매우 낮은 값을 나타낸다. 이와 같이, 잔사의 불용성 식이섬유 함량이 높은 것은 폴리페놀 추출 과정에서 수용성 식이섬유가 소실되거나 상등액으로 이행되기 때문이다. 또한, 카카오 콩 껍질의 추출 잔사의 탄수화물 함량(50 ∼ 60 중량%)이 높고 환원당 함량이 낮아서 잔사는 주로 고분자 물질이기 때문에 식이섬유는 이 잔사에 주로 존재한다. 단백질(33.43 ± 0.09 중량%) 및 조회분(18.03 ± 0.67 중량%)의 함량은 상등액에서 더 높은데, 상기된 바와 같이 에탄올로 폴리페놀을 추출하는 과정에서 단백질 및 염류 등의 수용성 물질이 상등액으로 이행되었기 때문이다. 따라서, 이러한 잔사 시료를 고식이 섬유원으로 이용하게 된 것이다.Insoluble dietary fiber content of the residue from which the polyphenol component is removed by ethanol extraction of the cacao bean husks of the cacao extract of the present invention is increased as compared to cacao bean husks. In contrast, the insoluble and water-soluble dietary fiber contents of the supernatant are very low, 0.2 to 0.4% by weight and 2 to 3% by weight, respectively. As such, the high insoluble fiber content of the residue is due to the loss of the water soluble fiber or the transition to the supernatant during the polyphenol extraction process. In addition, dietary fiber is mainly present in this residue because the carbohydrate content (50 to 60% by weight) of the extract residue of the cacao bean husk is high and the reducing sugar content is low so that the residue is mainly a polymer material. The content of protein (33.43 ± 0.09 wt%) and crude ash (18.03 ± 0.67 wt%) is higher in the supernatant, and as described above, water-soluble substances such as proteins and salts are transferred to the supernatant during the extraction of polyphenols with ethanol. Because Therefore, these residue samples are to be used as a source of dietary fiber.
또한, 카카오 콩 껍질의 불용성 식이섬유의 함량은 25 ∼ 45 중량%이고 수용성 식이섬유의 함량은 5 ∼ 15 중량%로서, 특히 불용성 식이섬유의 함량이 더 높다. 이 밖에, 카카오 열매 추출물의 불용성 식이섬유의 함량은 29 ∼ 33 중량%이고 수용성 식이섬유의 함량은 3 ∼ 5 중량%로 카카오 콩 껍질 보다 오히려 낮은 함량을 나타내지만, 이것도 불용성 식이섬유의 함량이 수용성 식이섬유의 함량보다 높아서 고식이섬유원으로 이용할 수 있다. 그러나, 카카오 열매 자체만으로서는 카카오 껍질보다 경제적이지 못하다. 또한, 카카오 매스 추출물의 불용성 식이섬유의 함량은 25 ∼ 35 중량%이고 수용성 식이섬유의 함량은 1.5 ∼ 3 중량%로서, 역시 불용성 식이섬유의 함량이 더 높다. 그러나, 상기에서 언급하였듯이 카카오 매스 자체만으로는 초콜렛의 원료로서의 사용은 적절하지만 고식이섬유원으로는 사용이 제한적이고 경제적 측면에서도 불리하다.In addition, the content of insoluble dietary fiber of cacao beans peel is 25 to 45% by weight and the content of water-soluble dietary fiber is 5 to 15% by weight, in particular, the content of insoluble fiber is higher. In addition, the content of insoluble dietary fiber of cacao extract is 29 to 33% by weight and the content of water-soluble dietary fiber is 3 to 5% by weight, which is lower than that of cacao beans, but this is also insoluble fiber content. It is higher than dietary fiber and can be used as a source of high dietary fiber. However, cacao alone is not as economical as cacao peel alone. In addition, the content of insoluble dietary fiber of cacao mass extract is 25 to 35% by weight and the content of water-soluble dietary fiber is 1.5 to 3% by weight, which is also higher in insoluble dietary fiber. However, as mentioned above, the use of chocolate as a raw material of the cacao mass alone is appropriate, but the use as a dietary fiber source is limited and economically disadvantageous.
한편, 본 발명의 카카오 추출물에 함유된 식이섬유는 높은 보수력, 보유능, 점도특성과 우수한 글루코오스 및 담즙산의 흡수 지연효과를 보이는 물리 화학적 성질을 나타낸다.On the other hand, the dietary fiber contained in the cacao extract of the present invention exhibits a high water holding capacity, retention capacity, viscosity characteristics and physicochemical properties showing excellent delayed absorption of glucose and bile acids.
본 발명의 카카오 콩 껍질 추출물, 카카오 열매 및 카카오 매스, 특히 카카오 콩 껍질에서의 추출물의 식이섬유의 보수력(Water holding capacity, WHC)은 시판용 펙틴의 약 8배보다도 높은 약 9배의 물을 흡수하여 매우 높은 보수력을 나타낸다. 반면, 카카오 껍질(cacao husk)과 카카오 열매 자체의 보수력은 각 추출물의 식이섬유 보수력의 약 1/2에 불과하다.Water holding capacity (WHC) of dietary fiber of cacao bean peel extract, cacao fruit and cacao mass, especially cacao bean peel of the present invention absorbs about 9 times higher water than about 8 times of commercial pectin Very high water holding power. On the other hand, cacao husk (cacao husk) and cacao fruit itself is only about 1/2 of the dietary fiber retention of each extract.
본 발명의 카카오 추출물에 함유된 식이섬유는 또 다른 물리적인 성질인 우수한 보유능(Oil holding capacity, OHC)을 갖는다. 본 발명의 카카오 추출물에서 식이섬유의 보유능은 보수력의 35 ∼ 70 중량%를 나타내고, 카카오 매스에서의 식이섬유의 경우는 보유능이 보수력의 약 60 ∼ 70 중량%나 되기 때문에 이것을 물과 기름이 함께 존재하는 가공식품에 사용하면 효율적인 흡착을 유도할 수 있다.The dietary fiber contained in the cacao extract of the present invention has an excellent oil holding capacity (OHC) which is another physical property. In the cacao extract of the present invention, the retention capacity of the dietary fiber represents 35 to 70% by weight of the water holding capacity, and in the case of the dietary fiber in the cacao mass, the holding capacity is about 60 to 70% by weight of the water holding power. When used in processed foods that can induce efficient adsorption.
또한, 본 발명의 카카오 추출물의 식이섬유는 점성 및 겔 형성능을 갖는데 소화관 내용물의 점도를 높여줌으로써 내용물의 소화관 이동 속도 및 흡수에 대해 영향을 준다. 특히, 놀랍게도 본 발명에 따른 카카오 콩 껍질에서 얻은 잔사 추출물의 식이섬유는 카카오 열매 식이섬유보다 다소 높은 점도를 나타내고 또 수용성 식이섬유보다는 불용성 식이섬유에서 높은 점도를 갖는 특징을 보인다.In addition, the dietary fiber of the cacao extract of the present invention has a viscosity and gel forming ability to increase the viscosity of the digestive tract contents affects the digestive tract movement speed and absorption of the contents. In particular, surprisingly, the dietary fiber of the residue extract obtained from the cacao bean husks according to the present invention exhibits a somewhat higher viscosity than the cacao fruit fiber and a higher viscosity in the insoluble fiber than the water-soluble fiber.
한편, 본 발명에 의한 식이섬유가 함유된 카카오 추출물에 대한 글루코오스 투과 저지율과 담즙산 투과 저지율의 생체밖(in-vitro) 시험을 통한 측정결과는 매우 우수한 결과를 나타낸다.On the other hand, the measurement results through the in-vitro test of glucose permeation inhibition rate and bile acid permeation inhibition rate for the cacao extract containing the dietary fiber according to the present invention shows very good results.
본 발명의 카카오 추출물인 잔사의 불용성 식이섬유와 수용성 식이섬유의 글루코오스 흡수 지연효과를 보면 각각 17 ∼ 20 % 및 25 ∼ 30 %로서 불용성 식이섬유보다 수용성 식이섬유의 함량이 더 높은데, 그 이유는 일반적으로 식이섬유는 시간이 경과함에 따라 보수력이 증가하고 겔(gel) 구조를 형성하여 물성 변화와 함께 글루코오스를 가두는 효과를 나타내어 점성이 높은 수용성 식이섬유의 경우 그 효과가 높게 나타나고 불용성 식이섬유는 낮게 나타나기 때문이다. 따라서, 본 발명의 카카오 추출물은 적어도 종래의 불용성 식이섬유인 시판용 α-셀룰로오스의 9 ∼ 11 %보다 더 높기 때문에 이 추출물이 고식이섬유 제품으로 활용하는데 매우 유용한 것이다. 그러나, 카카오 콩 껍질 자체의 불용성 식이섬유의 글루코오스 흡수 억제 지수는 15 ∼ 30 %이고 수용성 식이섬유의 글루코오스 흡수 억제지수는 10 ∼ 25 %로서, 불용성 식이섬유의 억제지수가 높다. 이것은 잔사의 경우 에탄올 추출로 인한 용매처리 효과로 글루코오스 흡수 지연효과가 달라져서 수용성 식이섬유의 억제지수가 높지만, 카카오 껍질은 이러한 추출과정을 거치지 않기 때문에 불용성 식이섬유의 억제지수가 높게 나타난다. 따라서, 카카오 콩 껍질 자체를 그대로 사용하기보다는 오히려 본원 발명에서와 같이 폴리페놀을 제거한 에탄올 추출 잔사가 더욱 유리한 결과를 나타낸다는 것을 보여준다. 그리고, 카카오 원료콩의 채취시기나 채취장소에 따라서도 결과가 달라진다.Glucose absorption delay effects of the insoluble fiber and water-soluble fiber of the residue of the cacao extract of the present invention are 17 to 20% and 25 to 30%, respectively, which are higher in water-soluble dietary fiber than insoluble fiber. As dietary fiber increases over time, water retention capacity increases and gel structure is formed to show the effect of confining glucose with changes in physical properties. Highly viscous water-soluble fiber shows high effect and low insoluble fiber. Because it appears. Therefore, the cacao extract of the present invention is at least higher than 9 to 11% of commercially available α-cellulose, which is a conventional insoluble dietary fiber, so that the extract is very useful for use as a high dietary fiber product. However, the glucose absorption inhibition index of insoluble dietary fiber of cacao beans peel itself is 15-30%, and the glucose absorption inhibition index of water-soluble dietary fiber is 10-25%, which is high in inhibiting dietary fiber. This result shows that the retardation effect of soluble dietary fiber is high due to the delayed effect of glucose absorption due to solvent treatment effect due to ethanol extraction, but the inhibition index of insoluble dietary fiber is high because cacao bark does not go through this extraction process. Thus, rather than using the cacao bean hull itself, it is shown that the ethanol extract residue from which the polyphenol was removed as in the present invention has more favorable results. The results also vary depending on the season and location of the cacao soybeans.
한편, 본 발명의 카카오 추출물인 잔사의 담즙산 흡수 억제지수는 불용성 식이섬유가 30 ∼ 35 %이고 수용성 식이섬유의 경우는 60 ∼ 65 %로서, 상기에서 설명한 바와 같이 수용성 식이섬유의 점도가 크기 때문에 함량이 더 높다. 그러므로, 본 발명의 카카오 추출물은 종래의 수용성 식이섬유인 시트러스 펙틴, 구아껌 등의 20 ∼ 32 %보다 답즙산 억제지수가 월등히 우수하다. 그리고 불용성 식이섬유의 담즙산 억제지수도 종래의 5 ∼ 10 %보다 우수하다. 또한, 카카오 콩 껍질의 불용성 식이섬유의 담즙산 흡수 억제지수는 35 ∼ 40 %이고 수용성 식이섬유의 경우는 55 ∼ 63 %로서 이것도 종래에 비해 함량이 더 높다. 이외에, 카카오 열매 및 카카오 매스의 답즙산 억제지수도 30 ∼ 35 %로 비슷한 경향을 나타낸다. 따라서, 수용성 식이섬유의 담즙산 억제지수가 카카오 열매 추출물을 제외한 나머지 추출물 모두 불용성 식이섬유의 2배에 가까운 값을 나타내므로 담즙산 흡수지연효과가 매우 뛰어남을 알 수 있다. 이와 같이, 불용성 식이섬유에 비해 수용성 식이섬유에서 그 효과가 높게 나타나는 것은 상기에서 설명한 바와 같이 수용성 식이섬유가 불용성 식이섬유에 비하여 점도가 높기 때문이다.On the other hand, the bile acid absorption inhibition index of the residue of the cacao extract of the present invention is 30 to 35% insoluble fiber and 60 to 65% for water-soluble dietary fiber, because the viscosity of the water-soluble dietary fiber is large as described above. Is higher. Therefore, the cacao extract of the present invention is much superior to the bile acid inhibition index than 20 to 32% of the conventional water-soluble dietary fiber citrus pectin, guar gum. In addition, the bile acid inhibition index of insoluble dietary fiber is also superior to that of the conventional 5-10%. In addition, the insoluble dietary fiber of cacao bean husks has a bile acid absorption inhibition index of 35 to 40% and a water-soluble dietary fiber of 55 to 63%, which is higher than the conventional one. In addition, the bile acid inhibition index of cacao fruit and cacao mass also shows a similar tendency of 30 to 35%. Therefore, it can be seen that the bile acid inhibitory index of the water-soluble dietary fiber exhibits almost twice the value of the insoluble dietary fiber except for the cacao extract, and thus the bile acid absorption delaying effect is excellent. As described above, the effect of the water-soluble dietary fiber is higher than that of the insoluble fiber, because the water-soluble dietary fiber has a higher viscosity than the insoluble fiber.
한편, 본 발명에 의한 카카오 추출물의 생체밖(In-vitro) 시험을 통한 생리활성을 보면 위에서부터 소장으로의 탄수화물 이동속도를 지연시키고 전분 및 다른 당류의 소화를 저해시키기 때문에 당질의 흡수를 감소시키며, 또 상피세포 표면으로 소화산물의 진입을 지연시키기 때문으로, 소장에서의 당 흡수를 지연시켜 혈당과 혈중 인슐린치를 감소시킨다. 또한, 본 발명에 따른 식이섬유가 함유된 카카오 추출물은 혈청의 중성지질, 인지질 및 콜레스테롤 농도를 현저하게 낮추어서 지질대사 불균형을 조절시키는 등 당뇨병과 성인병의 예방과 치료에 탁월한 효과를 보인다.On the other hand, the physiological activity of the cacao extract according to the present invention in vitro (in-vitro) to reduce the absorption of sugar because it delays the rate of carbohydrates from the stomach to the small intestine and inhibits the digestion of starch and other sugars In addition, because it delays the entry of digestive products to the surface of epithelial cells, it delays the absorption of glucose in the small intestine, thereby reducing blood sugar and insulin levels. In addition, cacao extract containing the dietary fiber according to the present invention shows an excellent effect in the prevention and treatment of diabetes and adult diseases, such as significantly lowering the level of neutral lipids, phospholipids and cholesterol of the serum to control the metabolic imbalance.
이하 본 발명을 실시예에 의거 더욱 상세히 설명하겠는바, 본 발명이 실시예에 의해 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited by Examples.
실시예 1 ∼ 4Examples 1-4
카카오 원료콩은 L-사가 구입하여 사용하는 가나(Ghana)산의 카카오 콩(Theobroma cacao L.)을 사용하였다. 그리고, 카카오 매스 가공의 최신 공법으로 알려진 니브(nib)를 균일한 작은 조각으로 만들어서 로우스팅(roasting) 처리하는 니브 로우스팅 과정(nib roasting process)인 도 1과 같은 L-사의 BTC(Better Taste and Color) 공정으로서, 카카오 원료콩을 0.8 bar의 압력과 170 ℃의 온도에서 열적 예열 처리한 다음 이것을 분쇄하고 선별하는 공정을 거친 후, 카카오 원료콩의 껍질을 탈각시키고 나서, 얻어진 카카오 콩 껍질에 에탄올을 첨가하여 1000 ∼ 3000 rpm의 원심분리기로 추출하여 잔사와 상등액으로 분획하여 잔사 추출물을 얻었다. 또한, 상기의 카카오 콩 껍질이 탈각되는 공정(winnowing)에서 분쇄 및 선별하여 껍질이 제거된 카카오 콩 분쇄물(winnowed nib)을 얻었고, 이 니브를 1.2 bar의 압력에서 10분 동안 반응(nib reaction) 처리한 후 카카오 니브(reacted nib)를 제조하였다. 그런 다음 132 ℃의 온도와 D.R.(Discharge Rate)이 1000 ㎏/hr의 조건에서 건조 및 볶음 처리후 갈아서 초콜렛의 주원료인 카카오 매스를 제조하였다.The cacao raw soybeans were used by the Ghana cacao beans (Theobroma cacao L.) purchased and used. In addition, a nib roasting process, which is a process known as the latest method of cacao mass processing, is made into a uniform small piece and roasted. As a color process, cacao beans are thermally preheated at a pressure of 0.8 bar and a temperature of 170 ° C., followed by a process of crushing and sorting the cacao beans. Was added and extracted with a centrifuge at 1000 ~ 3000 rpm and fractionated into a residue and a supernatant to obtain a residue extract. In addition, the cacao bean husks were pulverized and screened in a winnowing process to obtain shelled cacao soybean meal (winnowed nib) from which the nibs were reacted for 10 minutes at a pressure of 1.2 bar. After treatment, cacao nibs were prepared. Then, after drying and roasting at a temperature of 132 ° C. and D.R. (Discharge Rate) of 1000 kg / hr, the cacao mass, which is the main raw material of chocolate, was prepared.
실시예 5Example 5
상기 실시예 1에서 제조된 잔사 추출물에 대한 식이섬유 추출 및 분획은 도 2와 같은 프로스키(Proskey)법으로 실시하였다. 잔사를 30메쉬의 체에 통과시킨 후, 디에틸에테르를 사용하여 속스렛(Soxhlet) 법으로 12시간 동안 탈지하였다. 70 ℃로 유지한 건조기에서 건조한 다음 냄새를 제거하고 밀봉 후 냉장고에 보관하여 식이섬유 추출 시료로 사용하였다. 이렇게 준비된 잔사 시편 0.5g에 포스페이트 완충용액(0.08 M, pH 6, 25 ㎖)을 첨가하여 95 ∼ 100 ℃에서 30분 동안 테마밀(temamyl) 처리 후, 수산화나트륨(0.275 N, 5 ㎖)으로 pH를 7.5로 조절하고 60 ℃에서 30분 동안 프로테아제(protease) 처리하였다. 여기에 0.325 M의 염산 5 ㎖를 첨가하여 pH를 4 ∼ 4.6으로 조절하고 60 ℃에서 30분 동안 아밀로글루코시다제(amyloglucosidase) 처리를 한 다음 증류수로 세척 및 여과하여 식이섬유의 여과물과 잔사를 얻었다. 식이섬유 여과물에 60 ℃의 온도에서 에탄올(95%)을 첨가하고 이것을 다시 여과하는데, 이때 세척은 에탄올(78%, 95%) 및 아세톤으로 세척하고 건조 및 칭량하여 수용성 식이섬유를 얻었다. 또한, 상기의 식이섬유 잔사를 그대로 에탄올(95%)과 아세톤으로 세척하고 건조 및 칭량하여 불용성 식이섬유를 얻었고, 이 밖에 카카오 열매, 카카오 분쇄물, 카카오 니브 및 카카오 매스에 대해서도 동일한 방법으로 식이섬유를 얻었다.Dietary fiber extraction and fractionation of the residue extract prepared in Example 1 was carried out by the Proskey (Proskey) method as shown in FIG. The residue was passed through a 30 mesh sieve and then degreased using Soxhlet for 12 hours using diethyl ether. After drying in a dryer maintained at 70 ℃ was removed odor, sealed and stored in the refrigerator was used as a dietary fiber extract sample. To 0.5 g of the residue specimen thus prepared, phosphate buffer solution (0.08 M, pH 6, 25 ml) was added, followed by tetamyl treatment at 95-100 ° C. for 30 minutes, followed by pH with sodium hydroxide (0.275 N, 5 ml). Was adjusted to 7.5 and treated with protease at 60 ° C. for 30 minutes. To this, 5 ml of 0.325 M hydrochloric acid was added to adjust the pH to 4 to 4.6, treated with amyloglucosidase at 60 ° C. for 30 minutes, washed with distilled water, filtered and the filtrate and residue of dietary fiber. Got. Ethanol (95%) was added to the dietary fiber filtrate at a temperature of 60 ° C. and this was filtered again, wherein the wash was washed with ethanol (78%, 95%) and acetone, dried and weighed to obtain a water soluble fiber. In addition, the dietary fiber residue was washed with ethanol (95%) and acetone as it is, and dried and weighed to obtain insoluble dietary fiber. In addition, dietary fiber in the same way for cacao fruit, cacao grind, cacao nib and cacao mass. Got.
실시예 6Example 6
상기 실시예 1에서 제조된 카카오 콩 껍질의 수율은 12.63±1.38 중량%, 카카오 콩 분쇄물 86.77±1.52 중량%, 카카오 니브 95.95±0.49 중량%, 카카오 매스 84.18±0.23 중량%로, 각각에 용매를 첨가한 다음 혼합하여 추출물을 제조하였고, 용매를 제거하고 건조한 다음, 이에 대한 식이섬유 함유량 확인을 위해 상기 실시예 2와 같은 방법으로 식이섬유를 분획 및 추출하였다.Yield of cacao beans prepared in Example 1 is 12.63 ± 1.38% by weight, cacao beans ground 86.77 ± 1.52% by weight, cacao nibs 95.95 ± 0.49% by weight, cacao mass 84.18 ± 0.23% by weight, each solvent The mixture was added and then mixed to prepare an extract. The solvent was removed and dried, and then the fiber was fractionated and extracted in the same manner as in Example 2 to confirm the dietary fiber content thereof.
비교예 1 ∼ 3Comparative Examples 1 to 3
치커리, 알로에 베라 및 야콘은 통상적 시판되고 있는것을 구입하여 이것을 비교예로 하였으며, 상기 실시예 5의 프로스키 방법으로 식이섬유를 분획 및 추출하여 결과를 표 1에 나타내었다.Chicory, aloe vera and yacon were purchased commercially available as a comparative example, and the fiber was fractionated and extracted by the proski method of Example 5, the results are shown in Table 1.
실험예 1: 카카오 추출물의 식이섬유 함량과 구성성분의 조성Experimental Example 1: Dietary Fiber Content and Composition of Cacao Extracts
① 불용성 식이섬유(Insoluble dietary fiber, IDF)의 함량① Content of Insoluble dietary fiber (IDF)
상기에서 제조된 카카오 추출물의 불용성 식이섬유 함량은 프로스키(Prosky)법에 의하여 구하였다. 각각의 추출물을 2 개씩 0.5 g 칭량하여 AOAC(Association of Official Analytical Chemists)법에 의한 총 식이섬유 분석법과 동일한 방법으로 효소적 가수분해 과정을 거친 후, 용액(enzyme digest)을 미리 함량을 구한 B2 도가니 및 500 ㎖ 감압 플라스크에 감압 여과하고 다시 아세톤(200 ㎖, 2회)으로 세척하여 감압 여과하였다. 식이섬유가 들어있는 2개의 B2 도가니 중 하나는 회화시켜 회분 함량 (A)를 구하였고 또 다른 하나로 단백질 함량 (P)를 측정하였다. 이때 회분함량은 550 ℃의 직접회화법, 단백질 함량은 킬달(Kjeldahl: Kjeltec Auto 1035 Sampler System, Tecator, Sweden)법으로 측정하였다. 이러한 모든 과정을 블랭크(blank) 에서도 함께 진행하였고 이로부터 얻은 값(B)을 건조후의 무게에서 빼 주었으며 다음 수학식 1로부터 불용성 식이섬유의 함량을 계산하였고 결과를 표 1에 나타내었다.The insoluble dietary fiber content of the cacao extract prepared above was determined by the Prosky method. B2 crucible was weighed 0.5 g of each extract and subjected to enzymatic hydrolysis in the same way as the total fiber analysis by AOAC (Association of Official Analytical Chemists) method. And filtered under reduced pressure in a 500 mL vacuum flask, washed with acetone (200 mL, twice) and filtered under reduced pressure. One of the two B2 crucibles containing dietary fiber was incinerated to obtain ash content (A) and another to measure protein content (P). The ash content was measured by direct firing at 550 ° C. and the protein content by Kjeldahl: Kjeltec Auto 1035 Sampler System, Tecator, Sweden. All these processes were also carried out on a blank and the value obtained therefrom (B) was subtracted from the weight after drying. The content of insoluble dietary fiber was calculated from Equation 1 below and the results are shown in Table 1.
R1및 R2: 잔사(Residue) 무게, S1및 S2: 시료 무게R 1 and R 2 : Residue weight, S 1 and S 2 : Sample weight
P : IDF의 단백질 함량, A : IDF의 재 함량, B : 블랭크P: protein content of IDF, A: ash content of IDF, B: blank
② 수용성 식이섬유(Soluble dietary fiber, SDF)의 함량② The content of soluble dietary fiber (SDF)
상기에서 제조된 카카오 추출물의 수용성 식이섬유의 분석도 실험예 1과 같이 프로스키법에 의하여 다음과 같이 실시하였다. 즉, 불용성 식이섬유 분석시 보관해 두었던 500 ㎖ 감압 플라스크의 여과액을 증류수로 100 ㎖까지 맞추고 600 ㎖ 비이커에 옮겨 여기에 미리 60 ℃로 가열한 에탄올(95 중량%, 400 ㎖)를 가하고 실온에서 60 분간 정치시켜 수용성 식이섬유의 응집과 침전이 되도록 하였다. 미리 함량을 구한 2 개의 여과용 유리 도가니(1G3 도가니)에 정치 용액을 감압여과하고 20 ㎖의 에탄올(78 중량%)로 3회, 10 ㎖의 에탄올(95 중량%)로 2회, 20 ㎖의 아세톤으로 2회씩 세척하여 식이섬유가 들어있는 1G3 도가니를 실험예 1과 마찬가지로 하나는 회분 함량(A), 또다른 하나는 단백질 함량(P)을 구해 잔사의 무게에서 빼주었다. 이 모든 과정을 블랭크에서도 함께 진행하였으며 수용성 식이섬유 함량은 다음 수학 식 2로 계산하였고 결과를 표 1에 나타내었다.Analysis of the water-soluble dietary fiber of the cacao extract prepared above was carried out by the proski method as in Experimental Example 1. That is, the filtrate of the 500 ml vacuum flask stored in the analysis of insoluble fiber was adjusted to 100 ml with distilled water, and transferred to a 600 ml beaker, to which ethanol (95 wt%, 400 ml), which had been previously heated to 60 ° C., was added at room temperature. It was allowed to stand for 60 minutes to allow aggregation and precipitation of the water-soluble dietary fiber. The fixed solution was filtered under reduced pressure into two filtration glass crucibles (1G3 crucibles) prepared in advance, three times with 20 ml of ethanol (78 wt%), twice with 10 ml of ethanol (95 wt%), and 20 ml of After washing twice with acetone, 1G3 crucible containing dietary fiber was obtained in the same way as Experimental Example 1, one ash content (A) and the other protein content (P), and subtracted from the weight of the residue. All of these processes were also performed in the blank and the water soluble fiber content was calculated by the following Equation 2 and the results are shown in Table 1.
R1및 R2: 잔사(Residue) 무게, S1및 S2: 시료 무게R 1 and R 2 : Residue weight, S 1 and S 2 : Sample weight
P : SDF의 단백질 함량, A : SDF의 재 함량, B : 블랭크P: protein content of SDF, A: ash content of SDF, B: blank
③ 총 식이섬유(Total dietary fiber, TDF)의 함량③ The content of total dietary fiber (TDF)
상기에서 제조된 카카오 유래의 각 추출물에 대한 총 식이섬유 함량은 프로스키법으로 추출하여 불용성 식이섬유와 수용성 식이섬유 함량을 구한후 이들의 합(TDF, 중량% = IDF + SDF)으로 계산하였고 결과를 표 1에 나타내었다.The total dietary fiber content of each of the cacao-derived extracts prepared above was extracted by proski method, and the content of insoluble fiber and water-soluble dietary fiber was calculated and then calculated as the sum of these (TDF, weight% = IDF + SDF). Is shown in Table 1.
④ 불용성 식이섬유의 조성④ Composition of insoluble dietary fiber
상기에서 추출된 카카오 추출물의 불용성 식이섬유의 구성성분과 그 함량은 셀룰로오스, 헤미셀룰로오스 및 리그닌을 포함하는 중성 세정 섬유(NDF)와 셀룰로오스와 리그닌을 포함하는 산 세정 섬유(ADF)의 함량을 측정하여 구하였다. 즉, 셀룰로오스 함량은 ADF와 리그닌 함량의 차이로 구하였으며 헤미셀룰로오스의 함량은 NDF와 ADF의 함량의 차이로, 그리고 펙틴의 함량은 IDF와 NDF의 함량 차이로 다음 수학식 3과 같이 구하였다.Components of the insoluble dietary fiber and its content of the extracted cacao extract was measured by measuring the content of neutral cleansing fiber (NDF) containing cellulose, hemicellulose and lignin and acid cleansing fiber (ADF) containing cellulose and lignin It was. That is, the cellulose content was determined by the difference between the ADF and the lignin content, the hemicellulose content was calculated by the difference between the contents of the NDF and ADF, and the pectin content was calculated by the following equation (3).
헤미셀룰로오스 (중량%) = NDF - ADFHemicellulose (% by weight) = NDF-ADF
펙틴 (중량%) = TDF - NDFPectin (% by weight) = TDF-NDF
우선, NDF 함량을 측정을 위해 반 소에스트(Van Soest)와 윈(Wine)의 방법을 일부 변경하여 정량하였고, 몽게우(Mongeau)법에 따라 200 ㎖ 플라스크에 α-아밀라제(VI-B 형, A-3176 형, procine 췌장으로부터) 1.7g과 포스페이트 완충용액(0.1M, pH 7.0) 100 ㎖를 취한 후 15분 동안 교반하여 녹였고 4 ℃에서 1500 × g로 10분동안 원심분리하였다. 상등액을 4 ℃의 차가운 공간에서 1G3 도가니로 여과하고 α-아밀라제용액으로 세척하였고 사용 전까지 냉장 보관하였다. 50 ㎖ 플라스크에 카카오 추출물 시편 1g을 칭량하고 앞에서 조제한 α-아밀라제 용액 30 ㎖를 가하여 55 ℃의 진탕 항온수조에서 12시간 배양 한 다음, 500 ㎖ 환류 플라스크에 옮겼다. 중성 세정 용액(1ℓ 증류수에 소듐 라우릴 설페이트 30 g, 다이소듐 EDTA 18.61 g, 소듐 보레이트 데카하이드레이트 6.81 g, 다이소듐 하이드로겐 포스페이트(무수물) 4.56 g을 차례로 녹인 후 2-에톡시-에탄올 10 ㎖를 가하고 실온에서 H3PO4(인산) 용액으로 pH를 6.9 ∼ 7.1로 조절한 용액) 100 ㎖와 2 ㎖의 데카하이드로나프탈렌을 순차적으로 첨가하였다. 0.5 g의 소듐 설파이트를 이 용액에 완전히 녹인 다음 역류 냉각기를 달아 끓는 항온수조에서 60분간 가열하였고 1G3 도가니를 525 ℃ 회화로에서 1시간 동안 회화시켜 130 ℃의 건조오븐에서 3시간 동안 건조시키고 무게를 칭량하였다. 이 1G3 도가니에 가열시킨 용액을 감압여과하고 20 ㎖의 뜨거운 증류수로 2번 씻어내었다. NDF가 들어있는 1G3 도가니를 105 ℃의 건조오븐에서 12시간동안 건조시켜 건조기에서 방냉하여 칭량하고 이것을 525 ℃에서 3시간 동안 회화시키고 무게를 칭량하였다. 이 모든 과정을 블랭크에서도 함께 진행하였고 이 값을 건조후의 무게에서 빼주었으며 NDF 함량을 다음 수학식 4로부터 계산하였다.First, the NDF content was quantified by partially modifying the methods of Van Soest and Winine, and in a 200 ml flask according to Mongeau method, α-amylase (type VI-B, 1.7 g of A-3176, from procine pancreas) and 100 ml of phosphate buffer solution (0.1M, pH 7.0) were taken, dissolved for 15 minutes, and centrifuged at 1500 x g for 10 minutes at 4 ° C. The supernatant was filtered through a 1G3 crucible in a cold space at 4 ° C., washed with α-amylase solution and stored refrigerated until use. 1 g of cacao extract specimens were weighed into a 50 ml flask, 30 ml of the α-amylase solution prepared above was added thereto, and then incubated in a shake bath at 55 ° C. for 12 hours, and then transferred to a 500 ml reflux flask. Neutral washing solution (30 g of sodium lauryl sulfate, 18.61 g of disodium EDTA, 6.81 g of sodium borate decahydrate, 4.56 g of disodium hydrogen phosphate (anhydride) in 1 liter distilled water, and then 10 ml of 2-ethoxy-ethanol 100 ml and 2 ml of decahydronaphthalene were added sequentially at room temperature with a solution of pH adjusted to 6.9 to 7.1 with H 3 PO 4 (phosphate) solution. 0.5 g of sodium sulfite was completely dissolved in this solution, and then heated with a countercurrent cooler for 60 minutes in a boiling water bath. The 1G3 crucible was incubated for 1 hour in a 525 ° C incinerator and dried for 3 hours in a drying oven at 130 ° C. Was weighed. The solution heated in this 1G3 crucible was filtered under reduced pressure and washed twice with 20 ml of hot distilled water. The 1G3 crucible containing NDF was dried in a drying oven at 105 ° C. for 12 hours, cooled in a drier, weighed and weighed at 525 ° C. for 3 hours. All these processes were carried out together with the blanks, and this value was subtracted from the weight after drying and the NDF content was calculated from the following equation (4).
S : 시료의 무게, W1: 건조후의 무게S: weight of sample, W 1 : weight after drying
W2: 회화후의 무게, B : 블랭크의 무게W 2 : weight after painting, B: weight of blank
ADF의 함량은 AOAC법에 따라 구하였다. 500 ㎖의 환류 플라스크에 시료 1 g을 칭량하고 산 세정 용액(20 g의 메틸트리메틸 암모늄 브로마이드/ 1ℓ, 1 N 황산) 100 ㎖를 첨가한 후, 역류냉각기를 달아서 끓는 항온수조에서 60분간 가열하였다. 1G3 도가니를 525 ℃에서 1시간 동안 회화시키고, 105 ℃의 건조 오븐에서 3시간동안 건조시키고 방냉하여 무게를 칭량하였고, 이 1G3 도가니에 가열시킨 용액을 감압여과하고 뜨거운 증류수 100 ㎖ 정도를 사용하여 세정제를 완전히 씻어낸 후 20 ㎖의 아세톤으로 2회 씻어내었다. ADF가 들어있는 1G3 도가니를 105 ℃의 건조 오븐에서 12시간동안 건조시키고 건조기에서 방냉하였고 칭량 후 ADF의 함량을 다음 수학식 5에 따라 계산하였다.The content of ADF was determined by the AOAC method. 1 g of the sample was weighed into a 500 ml reflux flask, and 100 ml of an acid washing solution (20 g of methyltrimethyl ammonium bromide / 1 L, 1 N sulfuric acid) was added, followed by a countercurrent cooler and heated in a boiling water bath for 60 minutes. The 1G3 crucible was incubated for 1 hour at 525 ° C., dried for 3 hours in a drying oven at 105 ° C., cooled and weighed. The solution heated in the 1G3 crucible was filtered under reduced pressure and washed with about 100 ml of hot distilled water. Was washed thoroughly and washed twice with 20 ml of acetone. The 1G3 crucible containing ADF was dried in a drying oven at 105 ° C. for 12 hours, allowed to cool in a drier, and the content of ADF was calculated according to the following equation 5 after weighing.
S : 시료의 무게(g), W1: 도가니의 무게(g)S: weight of sample (g), W 1 : weight of crucible (g)
W2: 건조후의 무게(g)W 2 : Weight after drying (g)
리그닌의 함량은 AOAC법에 따라 분석하였다. 즉, 앞에서 구한 ADF가 들어있는 1G3 도가니를 50 ㎖의 비이커에 넣은 후, 잔사를 표면이 덮힐 정도로 72 중량%의 황산(15 ℃)를 1G3 도가니에 가해 유리봉으로 저으면서 균질화하고 여기에 72 %의 황산을 1G3 도가니의 2/3 정도까지 붓고, 20 ∼ 23 ℃에서 1시간마다 교반해 주면서 3시간 동안 정치하였다. 1G3 도가니에 정치한 용액을 감압 여과한 다음 100 ㎖ 정도의 뜨거운 증류수로 씻어내고 산을 완전히 제거하였다. 리그닌이 들어있는 도가니를 105 ℃의 건조 오븐에서 12 시간동안 건조시킨 다음, 건조기에서 방냉하고 칭량하였고, 다시 500 ℃에서 3시간 동안 회화 시켜 다시 무게를 칭량하였다. 이 모든 과정을 블랭크에서도 함께 진행하고 이 무게를 건조후의 무게에서 빼주었으며 리그닌 함량은 다음 수학식 6을 사용하여 계산하였다.The content of lignin was analyzed according to the AOAC method. In other words, put the 1G3 crucible containing the ADF obtained above into a 50 ml beaker, and then homogenize while adding 72% by weight of sulfuric acid (15 ° C) to the 1G3 crucible so that the residue covers the surface and stir it with a glass rod. Sulfuric acid was poured to about 2/3 of the 1G3 crucible and allowed to stand for 3 hours while stirring at 20-23 ° C for 1 hour. The solution left in the 1G3 crucible was filtered under reduced pressure, washed with hot distilled water of about 100 ml, and the acid was completely removed. The crucible containing lignin was dried in a drying oven at 105 ° C. for 12 hours, then cooled in a dryer and weighed, and again weighed by incubating at 500 ° C. for 3 hours. All of these processes were performed together with the blank, and the weight was subtracted from the weight after drying, and the lignin content was calculated using the following Equation 6.
S : 시료의 무게(g), W1: 72 중량%의 황산 처리 후 건조한 무게S: weight of sample (g), W 1 : dry weight after sulfuric acid treatment of 72 wt%
W2: 회화후의 무게, B : 블랭크의 무게W 2 : weight after painting, B: weight of blank
불용성 식이섬유의 ADF와 NDF의 함량을 상기의 수학식에 의해서 구하였고 그에 대한 결과를 표 2에 나타내었다.The contents of ADF and NDF of insoluble fiber were calculated by the above equations and the results are shown in Table 2.
상기의 방법으로 측정한 카카오 콩 껍질의 셀룰로오스 함량은 18.55 중량%, 헤미셀룰로오스 함량은 5.35 중량%, 리그닌의 함량은 17.75 중량%, 펙틴의 함량은 12.45 중량%이었다. 그리고, 카카오 콩 껍질의 추출잔사의 경우 셀룰로오스, 헤미셀룰로오스, 리그닌 및 펙틴의 함량은 각각 20.05, 4.57, 19.33 및 9.66 중량%로 카카오 콩 껍질과 비교하면 불용성 식이섬유의 증가로 각 성분 모두 다소 증가함을 보였다. 또한, 카카오 열매의 셀룰로오스, 헤미셀룰로오스, 리그닌 및 펙틴의 함량은 각각 9.26 중량%, 2 중량%, 19.25 중량% 및 2.84 중량% 이었다. 그리고, 카카오 분쇄물, 카카오 니브 및 카카오 매스의 셀룰로오스의 함량은 각각 7.27, 6.9 및 6.05 중량%이고, 헤미셀룰로오스의 함량은 각각 3.3, 2.25 및 3.75 중량%, 리그닌은 각각 20.33, 21.8 및 20.45 중량%, 펙틴은 각각 2.72, 2.02 및 0.53 중량%이었다. 상기의 결과에 따르면 본 발명의 추출물은 리그닌의 함량이 종래의 식이섬유 조성 함량인 곡류와 야채류의 1.98 및 0.29 중량% 보다 훨씬 더 많았다.The cellulose content of the cacao bean husk measured by the above method was 18.55 wt%, the hemicellulose content was 5.35 wt%, the lignin content was 17.75 wt%, and the pectin content was 12.45 wt%. In addition, the content of cellulose, hemicellulose, lignin and pectin was 20.05, 4.57, 19.33 and 9.66% by weight, respectively. Seemed. In addition, the contents of cellulose, hemicellulose, lignin and pectin of cacao were 9.26 wt%, 2 wt%, 19.25 wt% and 2.84 wt%, respectively. And, the content of cellulose of cacao crush, cacao nib and cacao mass is 7.27, 6.9 and 6.05 wt%, respectively, the content of hemicellulose is 3.3, 2.25 and 3.75 wt%, lignin 20.33, 21.8 and 20.45 wt%, respectively. Pectin was 2.72, 2.02 and 0.53 wt%, respectively. According to the above results, the extract of the present invention was much higher in lignin content than 1.98 and 0.29% by weight of the grains and vegetables of the conventional dietary fiber composition.
실험예 2: 카카오 식이섬유의 물리화학적 특성Experimental Example 2: Physicochemical Properties of Cacao Dietary Fiber
① 보수능 (Water holding capacity, WHC)① Water holding capacity (WHC)
소술스키와 카덴의 방법에 따라 카카오 콩 껍질, 카카오 열매 및 카카오 매스 추출물에서 추출한 식이섬유 시편 1 g을 50 ㎖ 원심분리용 셀에 넣고 증류수 20 ㎖를 첨가하여 10 분간 방치 후 유리막대로 10분 간격으로 30초 동안 3회 반복하여 저어서 10,000 rpm에서 25분간 원심분리하여 상등액을 제거한 후 잔사의 무게를 측정하였다. 카카오 콩 껍질, 카카오 열매 및 카카오 매스 추출물의 식이섬유 보수력은 각각 9.12, 9.20 및 3.52 g water retained/ g solid이었다.1 g of dietary fiber specimens extracted from cacao bean husk, cacao fruit and cacao mass extract were added to a 50 ml centrifuge cell and 20 ml of distilled water was added for 10 minutes. The mixture was stirred three times for 30 seconds, centrifuged at 10,000 rpm for 25 minutes to remove the supernatant, and then the residue was weighed. The fiber retention of cacao beans, cacao and cacao mass extracts was 9.12, 9.20 and 3.52 g water retained / g solid, respectively.
② 보유능 (Oil binding capacity, OBC)② Capacity (Oil binding capacity, OBC)
린 등의 방법에 따라 카카오 콩 껍질, 카카오 열매 및 카카오 매스 추출물에서 추출한 식이섬유 시편 1 g을 미리 함량을 구해놓은 원심분리용 셀에 넣고 콩기름(주, 해표) 5 ㎖를 첨가하여 5분 간격으로 30초씩 30분간 교반한 다음 10,000 rpm에서 25분간 원심분리하고 상등액을 제거한 후 무게를 측정하였다. 카카오 콩 껍질, 카카오 열매 및 카카오 매스 식이섬유의 보유능은 각각 3.30, 3.72 및 2.34 g oil retained/g solid로 보유능은 보수력의 36.2 ∼ 66.5 % 범위를 나타내었다.1 g of dietary fiber specimens extracted from cacao bean husks, cacao fruit and cacao mass extract were added to a centrifuge cell with a predetermined content, and 5 ml of soybean oil (note, haemul) was added at 30 minutes intervals. After stirring for 30 minutes each, centrifuged at 10,000 rpm for 25 minutes, the supernatant was removed and weighed. The retention capacities of cacao beans, cacao fruit and cacao mass fiber were 3.30, 3.72 and 2.34 g oil retained / g solid, respectively, and the retention capacity ranged from 36.2 to 66.5% of water holding capacity.
③ 점도③ viscosity
상기 실시예 1 ∼ 3에서 제조된 카카오 콩 껍질, 카카오 열매 및 카카오 매스 추출물의 식이섬유 그대로 또는 식이섬유에 0.1 ∼ 0.5 중량%의 안정제(펙틴)를 혼합하고 증류수에 현탁시켜 1 ∼ 3 중량% 현탁액으로 하였다. 60 ℃로 가온하고 다시 30 ℃로 냉각하여 회전 점도계(Brookfield DV-Ⅱ+, Brookfield Eng. labs Inc.) 또는 모세관 점도계(Cannon-Fenske, Cannon Instrument Co.)로 점도를 측정하였다. 1 %(w/v)의 카카오 콩 껍질 및 카카오 열매의 수용성 식이섬유의 점도는 1.363 및 1.339 mPaㆍS이고 점증제(0.1 % 펙틴 및 구아껌)를 첨가하면1.5배(1.902 ∼ 2.097 mPaㆍS) 및 2배(3.803 ∼ 3.858 mPaㆍS)의 점도를 나타내었고, 점증제 용액 중의 1 % 불용성 식이섬유 첨가 현탁액에 대한 점도를 측정한 결과, 0.1 % 펙틴 및 구아껌의 점도는 각각 1.57 및 3.16 mPaㆍS이었다. 0.1 % 펙틴 용액에서의 1 % 카카오 콩 껍질 및 카카오 열매의 식이섬유 점도는 4.47 및 3.97 mPaㆍS이고, 0.1 % 구아껌 용액에서의 1 % 카카오 콩 껍질 및 카카오 열매의 식이섬유 점도는 8.22 및 7.48 mPaㆍS이었고, 0.1 % 구아껌 용액중의 2 % 카카오 콩 껍질 및 카카오 열매에서는 50 rpm에서의 겉보기 점도값이 각각 21.76 및 17.01 mPaㆍS이었다.Dietary fiber of cacao bean husks, cacao fruit and cacao mass extract prepared in Examples 1 to 3 or 0.1 to 0.5% by weight of stabilizer (pectin) is mixed with dietary fiber and suspended in distilled water to 1 to 3% by weight suspension It was made. The viscosity was measured using a rotary viscometer (Brookfield DV-II +, Brookfield Eng. Labs Inc.) or a capillary viscometer (Cannon-Fenske, Cannon Instrument Co.) after warming to 60 ° C and cooling to 30 ° C. The viscosity of water-soluble dietary fiber of cacao beans and cacao beans at 1% (w / v) was 1.363 and 1.339 mPa · S, and 1.5 times (1.902 to 2.097 mPa · S) with the addition of thickeners (0.1% pectin and guar gum). ) And 2 times (3.803 to 3.858 mPa.S), and the viscosity of the 1% insoluble dietary fiber-added suspension in the thickener solution was measured. As a result, the viscosity of 0.1% pectin and guam was 1.57 and 3.16, respectively. mPa · S. The fiber viscosity of 1% cacao beans and cacao in 0.1% pectin solution was 4.47 and 3.97 mPa.S, and the fiber viscosity of 1% cacao beans and cacao in 0.1% guam gum was 8.22 and 7.48. mPa.S, and the apparent viscosity values at 50 rpm in 2% cacao bean husk and cacao fruit in 0.1% guam gum solution were 21.76 and 17.01 mPa.S, respectively.
실험예 3 : In-vitro법에 의한 글루코오스의 흡수 지연효과Experimental Example 3: Delayed Absorption Effect of Glucose by In-vitro Method
식이섬유가 투석막을 빠져나오는 글루코오스 및 담즙산의 투과를 지연하는 원리를 이용한 아디오톰레(Adiotomre) 등의 반투막 투과 실험법으로 측정하였다. 유리상태의 글루코오스는 투석막을 그대로 통과하지만 고분자 물질에 흡착된 글루코오스는 투석막을 통과하지 못하므로 투석 외액의 글루코오스 농도를 분석하여 측정하였다. 넓이 3.2 ㎠, 길이 10 ㎝의 투석막(Sigma D7884 : M.W. cut-off<1200)을 0.1 중량% 소듐 아자이드 용액에 하룻밤 담근 후 사용하며 투석막의 한쪽 끝을 목면실로 단단하게 묶은 후 투석막 내부에 카카오 추출물의 식이섬유 시편 0.2 g을 넣고 여기에 글루코오스 36 ㎎을 용해시킨 0.1 중량% 소듐 아자이드 용액 6 ㎖를 넣었다. 반대편의 투석막 끝도 단단히 묶은 후 150 ㎖ 용량의 용기에 넣고 14 시간동안 수화시켰으며 이때 대조구의 경우는 카카오 추출물의 식이섬유 시편만을 제외하여 마찬가지 방법으로 실시하고 수화 종료 후 용기에 0.1 중량% 소듐 아자이드 용액 100 ㎖를 첨가하였고 이를 37 ℃로 유지한 후 진탕 항온조에서 100 rpm으로 24시간동안 투과실험을 실시하며 일정시간(30분 ∼ 24 시간) 간격으로 투석외액 1 ㎖씩을 취하여 글루코오스 함량을 측정하였으며 흡수 지연 효과를 다음 식에 의해 계산하였다.The dietary fiber was measured by a semipermeable membrane permeation experiment such as Adiotomre using the principle of delaying the permeation of glucose and bile acids leaving the dialysis membrane. Glucose in the free state passes through the dialysis membrane as it is, but glucose adsorbed on the polymer material does not pass through the dialysis membrane. A dialysis membrane (Sigma D7884: MW cut-off <1200) of 3.2 cm2 in width and 10 cm in length is used after soaking in 0.1 wt% sodium azide solution overnight, and one end of the dialysis membrane is tightly bound with a cotton thread, and then the cacao extract inside the dialysis membrane. 0.2 g of dietary fiber specimens were added and 6 ml of a 0.1 wt% sodium azide solution in which 36 mg of glucose was dissolved. The dialysis membrane on the other side was also tightly bound and hydrated in a 150 ml container for 14 hours, except for the dietary fiber specimens of cacao extract, and 0.1 wt% sodium azide in the container after hydration. 100 ml of solution was added and maintained at 37 ° C. Permeation experiment was carried out for 24 hours at 100 rpm in shaking thermostat. Glucose content was measured by taking 1 ml of dialysis external solution at regular intervals (30 minutes to 24 hours). The delay effect was calculated by the following equation.
이때 글루코오스 함량은 ABTS 법에 따라 측정하는데, 0.1 중량% 소듐 아자이드 용액에 용해시킨 카카오 추출물의 식이섬유 시편 및 표준 글루코오스 용액 1 ㎖에 ABTS 시약(글루코오스 옥시다아제 60 ㎎, 퍼옥시다아제 6 ㎎ 및 ABTS 50 ㎎을 0.12 M 포스페이트 완충 용액 250 ㎖에 용해한 것) 5㎖를 첨가한 후, 실온에서 30 ∼ 40분간 방치하였고 450 nm에서의 흡광도를 측정하였다. 글루코오스의 경시변화와 흡수 억제지수를 구하여, 그 결과를 도 4a, 4b 및 표 3에 나타내었다.At this time, the glucose content was measured according to the ABTS method, and the ABTS reagent (60 mg of glucose oxidase, 6 mg of peroxidase and 50 mg of ABTS) was added to 1 ml of dietary fiber specimen and standard glucose solution of cacao extract dissolved in 0.1 wt% sodium azide solution. Was dissolved in 250 ml of 0.12 M phosphate buffer solution), and then allowed to stand at room temperature for 30 to 40 minutes, and the absorbance at 450 nm was measured. The change in glucose over time and the absorption inhibition index were obtained, and the results are shown in FIGS. 4A, 4B and Table 3.
실험예 4 : In-vitro법에 의한 담즙산의 흡수 지연효과Experimental Example 4: Effect of delaying absorption of bile acids by in-vitro method
담즙산 흡수 지연 효과는 글루코오스와 마찬가지로 유리상태의 담즙산이 투석막을 빠져나오므로 투석막 내부에 카카오 추출물의 식이섬유 시편 0.2 g을 넣고 0.1 중량% 소듐 아자이드 용액으로 조제한 0.05 M 포스페이트 완충용액(pH 7.0)에 1ℓ당 15 mmol 타우로콜린산(Sigma T ∼4009)을 녹인 용액 6 ㎖를 넣어 투석막 끝을 단단히 묶었다. 이것을 150 ㎖ 용량의 용기에 넣어 14시간 동안 수화시켰으며 대조구의 경우는 카카오 추출물 시편만을 제외하여 마찬가지로 실시하였다. 여기에 0.1 중량% 소듐 아자이드 용액으로 조제한 0.05 M 포스페이트 완충용액(pH 7.0) 100 ㎖를 첨가한 후, 37 ℃의 항온 수조에서 100 rpm으로 72시간 동안 진탕하였다, 일정시간 간격으로 1 ㎖씩을 취하여 담즙산의 함량을 측정하였으며 담즙산 흡수 지연 효과는 다음 식에 의해 계산하였다.The effect of delaying bile acid absorption is that, in the same way as glucose, free bile acid escapes the dialysis membrane, so that 0.2 g of dietary fiber specimens of cacao extract are added to the dialysis membrane in 0.05 M phosphate buffer (pH 7.0) prepared with 0.1 wt% sodium azide solution. 6 ml of a solution in which 15 mmol taurocholic acid (Sigma T-4009) was dissolved per liter was added, and the dialysis membrane tip was tightly bound. This was hydrated for 14 hours in a 150 ml container, and the control was similarly performed except for cacao extract specimens. 100 ml of 0.05 M phosphate buffer (pH 7.0) prepared with 0.1 wt% sodium azide solution was added thereto, followed by shaking for 72 hours at 100 rpm in a constant temperature water bath at 37 ° C. The content of bile acid was measured, and the effect of delaying bile acid absorption was calculated by the following equation.
이때 담즙산 함량은 보이드(Boyd) 등의 방법에 따라 측정하였다. 즉 담즙산(Sigma B-8756) 용액 1 ㎖에 70중량% 황산 용액 5 ㎖를 넣고 5분 후에 0.25 중량% 푸르푸랄(furfural) 용액 1 ㎖를 각각 첨가하였다. 60분간 방치하여 분홍색이 최대로 발색된 후의 최대 흡수 파장인 510 nm에서 흡광도를 측정하였다. 투석막을 통해 타우로콜린산의 양을 72 시간동안 경시적으로 측정하여 도 5a 및 도 5b에 나타내었다. 또한, 담즙산의 흡수 지연효과를 보다 명확히 알아보기 위하여 글루코오스에서와 마찬가지로 담즙산 흡수 억제지수(bile acid retardation index, %)를 구하였으며 그 결과를 표 4에 나타내었다.The bile acid content was measured according to the method of Boyd (Boyd) and the like. That is, 5 ml of a 70% by weight sulfuric acid solution was added to 1 ml of a bile acid (Sigma B-8756) solution, and 5 minutes later, 1 ml of 0.25% by weight furfural solution was added. After leaving for 60 minutes, the absorbance was measured at 510 nm, the maximum absorption wavelength after the maximum color development of pink. The amount of taurocholine acid through the dialysis membrane was measured over time for 72 hours and is shown in FIGS. 5A and 5B. In addition, the bile acid retardation index (bile acid retardation index,%) was obtained in the same manner as in glucose in order to clarify the effect of bile acid absorption.
실험예 5: 식이섬유가 함유된 카카오 추출물의 당 치료 효과 실험Experimental Example 5 Experiment of Sugar Treatment Effect of Cacao Extract Containing Dietary Fiber
① 실험동물을 통한 식이 및 물의 섭취량, 체중 및 배변량의 변화① Changes in diet, water intake, weight, and bowel movements through experimental animals
카카오 추출물의 식이섬유의 혈중 포도당 수준 및 지질대사 개선효과 등의 생리 효과를 조사하기 위하여 스트렙토조토신(STZ, Sigma S-0130)으로 당뇨를 유발시킨 당뇨쥐를 대상으로 다음과 같은 동물실험을 실시하였다. 실험동물은 5 주령의 수컷 S.D.(Sprague Dawley)계열로 대한 동물실험 센터로부터 분양 받아서 사용하였고, 실험식이의 조제로 동물실험에 사용한 표준 식이조성(중량%)은 표 5와 같으며 실험동물의 식이, 물의 섭취량, 체중, 배변량은 표 6과 같이 실시하였다.To investigate the physiological effects of cacao extracts, such as improving blood glucose levels and lipid metabolism, the following animal experiments were conducted in diabetic rats that induced diabetes with streptozotocin (STZ, Sigma S-0130). It was. The experimental animals were used from the animal testing center for the male SD (Sprague Dawley) family of 5 weeks old, and the standard diet composition (% by weight) used for the animal experiments as the preparation of the experimental diet is shown in Table 5. , Water intake, weight, defecation was performed as shown in Table 6.
실험동물을 온도 20 ∼ 22℃, 습도 50±10 %의 12시간 주기의 명암하에서 사육하였고 스테인레스 스틸 케이지에서 1주일 동안 고형사료로 적응시킨 후 당뇨를 유발시켰다. 당뇨 유발은 스트렙토조토신을 시트레이트 완충용액(pH 4.5)에 녹여 50 ㎎/㎏의 농도로 1회 복강 주사하고 12시간 동안 절식시킨 후, 꼬리 정맥에서 채혈하여 혈당 농도가 180 ㎎/㎗ 이상인 것들만 당뇨쥐로 사용하였고, 대조군은 0.01 M의 시트레이트 완충용액만을 사용하여 당뇨 유발군과 같은 방법으로 주사하였다. 실험동물은 적응사육 2 주후 정상쥐 표준식이군, 당뇨쥐 표준 식이균, 잔사군, 상등액군, 카카오 매스(카카오 매스) 군, 카카오 매스-NDF군으로 나누어 다시 2 주간 실험 식이로 사용하였으며 이때 각군은 체중이 비슷한 8마리를 1군으로 하였다. 물은 증류수를 사용하였고 식이는 자유로이 섭취하도록 하였으며 식이 섭취량, 변량 및 물 섭취량은 매 1일, 체중은 2일 간격으로 측정하였다.The animals were bred under a 12-hour cycle with a temperature of 20-22 ° C and a humidity of 50 ± 10%, and adapted to solid feed in a stainless steel cage for 1 week before inducing diabetes. Diabetes induction was performed by dissolving streptozotocin in citrate buffer (pH 4.5) once intraperitoneally at a concentration of 50 mg / kg, fasting for 12 hours, and collecting blood from the tail vein to achieve blood glucose levels above 180 mg / dl. Mice were used, and the control group was injected in the same manner as the diabetic induction group using only 0.01 M citrate buffer. After 2 weeks of adaptation, the experimental animals were divided into two groups: normal rat standard diet, diabetic rat standard diet, residue group, supernatant group, cacao mass (cacao mass) group, and cacao mass-NDF group. 8 animals of similar weight were used as the first group. Distilled water was used for water, and diet was freely ingested. Dietary intake, variance and water intake were measured every 1 day and body weight was measured every 2 days.
② 혈중 당농도 및 총 당화 혈색소 측정② Measurement of blood glucose level and total glycated hemoglobin
2주간의 실험식이 사육이 끝난 실험동물은 12 시간동안 절식시킨 후 에틸 에테르로 가볍게 마취시켜서 복부를 절개하고 심장에서 체혈하여 전혈을 얻었으며 이를 당화 혈색소 분석의 시편으로 하였다. 전혈은 항 응고제(heparin)가 들어있는 마이크로 헤마토크리트 튜브(GRAF Cat. No. 5.530-06)를 통과시킨 후 3000 rpm(4℃)에서 10분간 원심분리하여 혈청을 분리하였고, 혈당 및 지질 분석의 시편으로 하였다. 한편, 간장은 적출 후 살린(saline)으로 처리하였고, 여과지(Watman No. 2)로 수분을 제거한 다음 무게를 측정하였다. 이상의 시편들은 분석 전까지 -70 ℃의 냉동고에 보관하였고, 총 당화 혈색소를 측정하여 표 7 및 도 6에 나타내었다.After 2 weeks of experimental breeding, the animals were fasted for 12 hours, and then lightly anesthetized with ethyl ether to incise the abdomen and bleed from the heart to obtain whole blood, which was used as a specimen of glycated hemoglobin analysis. Whole blood was passed through a micro hematocrit tube containing anticoagulant (GRAF Cat. No. 5.530-06), and then serum was separated by centrifugation at 3000 rpm (4 ° C) for 10 minutes. It was made. On the other hand, soy sauce was extracted and treated with saline, water was removed with filter paper (Watman No. 2), and then weighed. The specimens were stored in a freezer at −70 ° C. until analysis, and the total glycated hemoglobin was measured and shown in Table 7 and FIG. 6.
혈당 측정은 혈청을 분리하고 GOD(글루코스-옥시다제) 측정법에 따라 조제된 아산제약의 혈당 측정용 킷(kit)(AM 201-K)를 사용하여 500 nm에서 비색 정량하였다.Blood glucose measurements were colorimetrically quantitated at 500 nm using a blood glucose measurement kit (AM 201-K) of Asan Pharmaceuticals prepared according to GOD (glucose-oxidase) assay.
③ 중성지질(triglyceride, TG)과 인지질(phospholipid)의 농도③ Concentrations of triglyceride (TG) and phospholipid
중성 지질 및 인지질의 측정은 혈청을 분리하고 효소 비색법을 이용한 아산제약의 킷(AM 1575-K) 및 이아트론 킷(Iatron kit)(PL-E(OM))을 사용하여 550 및 500 nm에서 각각 비색정량을 측정하였다.Determination of neutral lipids and phospholipids was performed at 550 and 500 nm using a kit of Asan Pharmaceuticals (AM 1575-K) and Iatron kit (PL-E (OM)) using serum separation and enzyme colorimetry, respectively. The colorimetric quantity was measured.
카카오 추출물의 식이섬유가 당뇨 동물에 대한 혈청내의 중성지질(triglyceride, TG)과 인지질(phospholipid)의 농도에 미치는 효과를 표 8에 나타내었다.Table 8 shows the effect of dietary fiber of cacao extract on the concentrations of triglyceride (TG) and phospholipid in serum of diabetic animals.
④ 콜레스테롤 함량④ cholesterol content
총 콜레스테롤 및 고밀도 지질 단백질(HDL, High-Density Lipoprotein)-콜레스테롤의 측정은 혈청을 분리하고 효소 비색법을 이용한 아산제약의 킷(콜레스테롤-효소 및 AM 203-K)을 사용하여 500 nm에서 각각 비색정량을 측정하였고, 저밀도 지질 단백질(LDL, Low-Density Lipoprotein)-콜레스테롤은 다음의 프리데왈드 식으로 구하였으며 동맥 경화지수(Atherogenic Index, AI)는 혈청 총-콜레스테롤에서 HDL-콜레스테롤을 뺀 수치를 HDL-콜레스테롤 값으로 나누어 산출하였다.Determination of total cholesterol and high-density lipoprotein (HDL) -cholesterol was performed by colorimetric determination at 500 nm, respectively, using serum kits and Asan Pharmaceutical's kit (cholesterol-enzyme and AM 203-K) using enzymatic colorimetry. Low-density lipoprotein (LDL) -cholesterol was measured by the following Friedelwald equation, and the atherosclerosis index (AI) was determined by subtracting HDL-cholesterol from serum total-cholesterol. Calculated by dividing by the cholesterol value.
STZ 투여 2주 후 실험동물 혈액중의 총 콜레스테롤, HDL-콜레스테롤, LDL-콜레스테롤 및 동맥 경화 지표에 미치는 영향을 측정하여 표 9에 나타내었다.Two weeks after STZ administration, the effects on total cholesterol, HDL-cholesterol, LDL-cholesterol, and atherosclerosis in experimental blood were measured and shown in Table 9.
본 동물실험에서 얻어진 측정결과의 통계적 유의성을 조사하기 위하여 SAS(Statistical Analysis System)프로그램을 이용하였으며, p<0.05 수준에서 던칸스 다중 시험(Duncan's multiple test)을 행하여 각 실험군별 평균치간의 유의성을 검정하였다.In order to investigate the statistical significance of the measurement results obtained in this animal experiment, SAS (Statistical Analysis System) program was used and Duncan's multiple test was performed at p <0.05 level to test the significance between the mean values of each experimental group. .
상술한 바와 같이, 본 발명에 따른 카카오 추출물은 종래의 카카오 매스 가공 중 전량 폐기되었던 카카오 콩 껍질을 이용함으로써, 비용절감과 환경 친화적인 효과가 있어 경제적이고, 특히 본 발명에서 얻은 카카오 콩 껍질의 추출 잔사는 식이섬유가 과량 함유되어 있고, 또한 식이섬유의 조성에서 리그닌의 함량이 풍부하여 우수한 글루코오스 및 담즙산 흡수 지연 효과와 혈당저하 및 지질대사개선효과를 나타내므로 당뇨병의 치료에 유용한 카카오 추출물의 기능성 제품화를 도모할 수 있는 효과가 있다.As described above, the cacao extract according to the present invention is economical by using the cacao bean husk, which has been discarded entirely during the processing of cacao mass, cost-effective and environmentally friendly, in particular the extraction of cacao bean husk obtained in the present invention The residue contains excessive amounts of dietary fiber, and also has abundant lignin content in the composition of dietary fiber, which shows excellent effect of delaying glucose and bile acid absorption, lowering blood sugar and improving lipid metabolism. There is an effect that can be planned.
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EP99941851A EP1168935A1 (en) | 1999-04-15 | 1999-09-03 | Cacao extract including dietary fiber |
JP2000611775A JP3533181B2 (en) | 1999-04-15 | 1999-09-03 | Cocoa extract containing dietary fiber |
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KR101189141B1 (en) * | 2009-01-30 | 2012-10-10 | 주식회사김정문알로에 | Method for preparing fiber of Aloe vera Linne parenchyma tissue for the dietary fiber |
KR20180063764A (en) | 2016-12-02 | 2018-06-12 | (주)나눔랩 | Shell method of cacao bean and foodstuffs comprising shelled cacao bean |
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KR100460103B1 (en) * | 2001-10-18 | 2004-12-04 | 롯데제과주식회사 | Drug with inhibitor effects on digestive system carcinogenesis containing cacao bean and husk fraction extract |
PL1728434T3 (en) | 2005-05-31 | 2009-05-29 | Kraft Foods R & D Inc | Process for the manufacture of a polyphenol-enriched composition from cocoa shell extraction |
JP5248755B2 (en) * | 2006-07-14 | 2013-07-31 | 株式会社ロッテ | Angiotensin I converting enzyme inhibitor and method for producing the same |
JP4941666B2 (en) * | 2007-10-19 | 2012-05-30 | 松谷化学工業株式会社 | Dietary fiber fortifier and food |
KR100888492B1 (en) * | 2008-07-31 | 2009-03-12 | 한국식품연구원 | Preparation method of dietary fiber from brewer's spent grain |
CN102697061A (en) * | 2012-06-04 | 2012-10-03 | 江苏南方食品科技有限公司 | Method for preparing soybean residue water soluble dietary fiber through microwave modification |
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JP6862219B2 (en) * | 2017-03-02 | 2021-04-21 | 昭和産業株式会社 | Confectionery mix, confectionery batter, confectionery and method of manufacturing confectionery |
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